STUDY QUESTION Can fluorescence lifetime imaging microscopy (FLIM) detect associations between the metabolic state of cumulus cell (CC) samples and the clinical outcome of the corresponding embryos? SUMMARY ANSWER FLIM can detect significant variations in the metabolism of CC associated with the corresponding embryos that resulted in a clinical pregnancy versus those that did not. WHAT IS KNOWN ALREADY CC and oocyte metabolic cooperativity are known to be necessary for the acquisition of developmental competence. However, reliable CC biomarkers that reflect oocyte viability and embryo developmental competency have yet to be established. Quantitative measures of CC metabolism could be used to aid in the evaluation of oocyte and embryo quality in ART. STUDY DESIGN, SIZE, DURATION A prospective observational study was carried out. In total, 223 patients undergoing IVF with either conventional insemination or ICSI at a tertiary care center from February 2018 to May 2020 were included, with no exclusion criteria applied. PARTICIPANTS/MATERIALS, SETTING, METHODS This cohort had a mean maternal age of 36.5 ± 4.4 years and an average oocyte yield of 16.9 (range 1–50). One to four CC clusters from each patient were collected after oocyte retrieval and vitrified. CC metabolic state was assessed using FLIM to measure the autofluorescence of the molecules NAD(P)H and FAD+, which are essential for multiple metabolic pathways. CC clusters were tracked with their corresponding oocytes and associated embryos. Patient age, Day 3 and Day 5/6 embryo morphological grades, and clinical outcomes of embryos with traceable fate were recorded. Nine FLIM quantitative parameters were obtained for each CC cluster. We investigated associations between the FLIM parameters and patient maternal age, embryo morphological rank, ploidy, and clinical outcome, where false discovery rate P-values of <0.05 were considered statistically significant. MAIN RESULTS AND THE ROLE OF CHANCE A total of 851 CC clusters from 851 cumulus–oocyte complexes from 223 patients were collected. Of these CC clusters, 623 were imaged using FLIM. None of the measured CC FLIM parameters were correlated with Day 3 morphological rank or ploidy of the corresponding embryos, but FAD+ FLIM parameters were significantly associated with morphological rank of blastocysts. There were significant differences for FAD+ FLIM parameters (FAD+ fraction engaged and short lifetime) from CC clusters linked with embryos resulting in a clinical pregnancy compared with those that did not, as well as for CC clusters associated with embryos that resulted in a live birth compared those that did not. LIMITATIONS, REASONS FOR CAUTION Our data are based on a relatively low number of traceable embryos from an older patient population. Additionally, we only assessed CCs from 1 to 4 oocytes from each patient. Future work in a younger patient population with a larger number of traceable embryos, as well as measuring the metabolic state of CCs from all oocytes from each patient, would provide a better understanding of the potential utility of this technology for oocyte/embryo selection. WIDER IMPLICATIONS OF THE FINDINGS Metabolic imaging via FLIM is able to detect CC metabolic associations with maternal age and detects variations in the metabolism of CCs associated with oocytes leading to embryos that result in a clinical pregnancy and a live birth versus those that do not. Our findings suggest that FLIM of CCs may be used as a new approach to aid in the assessment of oocyte and embryo developmental competence in clinical ART. STUDY FUNDING/COMPETING INTEREST(S) National Institutes of Health grant NIH R01HD092550-03 (to C.R., and D.J.N.). Becker and Hickl GmbH and Boston Electronics sponsored research with the loaning of equipment for FLIM. D.J.N. and C.R. are inventors on patent US20170039415A1. TRIAL REGISTRATION NUMBER N/A.

中文翻译：

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人类卵丘细胞的代谢成像揭示了与妊娠和活产的关联

研究问题 荧光寿命成像显微镜 (FLIM) 能否检测卵丘细胞 (CC) 样本的代谢状态与相应胚胎的临床结果之间的关联？摘要答案 FLIM 可以检测与导致临床妊娠的相应胚胎和未导致临床妊娠的胚胎相关的 CC 代谢的显着变化。已知的情况 已知 CC 和卵母细胞代谢协同对于获得发育能力是必要的。然而，反映卵母细胞活力和胚胎发育能力的可靠 CC 生物标志物尚未建立。 CC 代谢的定量测量可用于帮助评估 ART 中的卵母细胞和胚胎质量。研究设计、规模、持续时间进行了一项前瞻性观察研究。总共纳入了 223 名 2018 年 2 月至 2020 年 5 月期间在三级护理中心接受 IVF 常规授精或 ICSI 的患者，未采用排除标准。参与者/材料、背景、方法 该队列的母亲平均年龄为 36.5 ± 4.4 岁，平均卵母细胞产量为 16.9（范围 1-50）。取卵并玻璃化冷冻后，从每位患者收集一到四个 CC 簇。使用 FLIM 测量分子 NAD(P)H 和 FAD+ 的自发荧光来评估 CC 代谢状态，这对于多种代谢途径至关重要。 CC 簇与其相应的卵母细胞和相关胚胎一起被追踪。记录患者年龄、第 3 天和第 5/6 天胚胎形态分级以及具有可追踪命运的胚胎的临床结果。每个 CC 簇获得了 9 个 FLIM 定量参数。我们研究了 FLIM 参数与患者母亲年龄、胚胎形态等级、倍性和临床结果之间的关联，其中 <0.05 的错误发现率 P 值被认为具有统计显着性。主要结果和机会的作用 总共收集了来自 223 名患者的 851 个卵丘-卵母细胞复合体的 851 个 CC 簇。在这些 CC 簇中，有 623 个是使用 FLIM 成像的。测量的 CC FLIM 参数均与相应胚胎的第 3 天形态等级或倍性无关，但 FAD+ FLIM 参数与囊胚的形态等级显着相关。与那些没有临床妊娠的胚胎相关的CC簇相比，与导致临床妊娠的胚胎相关的CC簇的FAD+ FLIM参数（FAD+参与分数和寿命短）存在显着差异，与导致活产的胚胎相关的CC簇也存在显着差异。那些没有的。局限性和注意原因我们的数据基于来自老年患者群体的相对较少数量的可追踪胚胎。此外，我们仅评估了每位患者 1 至 4 个卵母细胞的 CC。未来在具有大量可追踪胚胎的年轻患者群体中开展工作，以及测量每位患者所有卵母细胞 CC 的代谢状态，将有助于更好地了解该技术在卵母细胞/胚胎选择方面的潜在效用。研究结果的更广泛意义 通过 FLIM 的代谢成像能够检测 CC 代谢与母亲年龄的关联，并检测与卵母细胞相关的 CC 代谢变化，这些卵母细胞导致临床妊娠和活产的胚胎与那些没有临床妊娠和活产的胚胎相关。我们的研究结果表明，CC 的 FLIM 可以作为一种新方法来帮助评估临床 ART 中的卵母细胞和胚胎发育能力。研究经费/竞争利益 美国国立卫生研究院授予 NIH R01HD092550-03（授予 CR 和 DJN）。 Becker and Hickl GmbH 和 Boston Electronics 通过向 FLIM 借用设备来赞助研究。 DJN 和 CR 是专利 US20170039415A1 的发明人。试用注册号 不适用。